[Mechanism of gypenoside LI inducing apoptosis and autophagy in clear cell renal cell carcinoma cells via ROS/JNK and Akt/mTOR signaling pathways].

This study aims to investigate the anti-proliferative effect and molecular mechanism of gypenoside LI(Gyp LI) in clear cell renal cell carcinoma(ccRCC) and elucidate its regulatory effect on apoptosis, autophagy, reactive oxygen species(ROS) signals, and related pathways. The effect of Gyp LI on ccRCC cell viability, proliferation, apoptosis, autophagy, and migratory capability was observed and assessed by cell counting kit-8(CCK-8), colony formation test, flow cytometry, Hoechst staining, wound healing assay, and transmission electron microscope(TEM). RNA sequencing, gene set enrichment analysis(GSEA), and Western blot were used to identify and verify the signaling pathways involved. The results show that Gyp LI significantly inhibits the proliferation of ACHN cells and 769-P cells, with IC_(50) values of 72. 30 and 62. 57 μmol·L~(-1) after treatment for 48 hours. Meanwhile, Gyp LI induced nuclear pyknosis and karyorrhexis, inhibited colony formation and cell migration, upregulated the expressions of B-cell lymphoma-2 associated X protein(Bax), cysteinyl aspartate-specific proteinase-3(caspase-3), microtubule-associated protein 1A/1B-light chain 3(LC3)-Ⅱ/LC3-Ⅰ, autophagy-related protein 5( ATG5), phosphorylated c-Jun n-terminal kinase( p-JNK), and phosphorylated p38 mitogen-activated protein kinase( p-p38 MAPK), downregulating the expressions of B-cell lymphoma-2( Bcl-2), sequestosome 1(p62), phosphorylated protein kinase B(p-Akt), and phosphorylated mammalian target of rapamycin(p-mTOR). Mechanistically, Gyp LI activated c-Jun N-terminal kinase( JNK)/p38 mitogen-activated protein kinase( p38 MAPK) pathways, inhibited protein kinase B(Akt)/mammalian target of rapamycin(mTOR) pathways, and promoted autophagy in ccRCC cells. Autophagy inhibition further enhanced apoptosis. Gyp LI also markedly induced ROS production, and this effect was partially reversed by N-acetyl-Lcysteine(NAC). Subcutaneous xenograft experiments of ACHN cells in BALB/c nude mice demonstrate that gypenosides can safely and effectively inhibit tumor growth. In conclusion, Gyp LI exerts anti-ccRCC effects by coordinately regulating ROS-mediated stress signaling, mitochondrial apoptosis, and autophagy.